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Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences

Mitochondrial matrix proteins synthesized in the cytosol often contain amino (N)-terminal targeting sequences (NTSs), or alternately internal targeting sequences (ITSs), which enable them to be properly translocated to the organelle. Such sequences are also required for proteins targeted to mitochon...

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Autores principales: Santos, Herbert J., Chiba, Yoko, Makiuchi, Takashi, Arakawa, Saki, Murakami, Yoshitaka, Tomii, Kentaro, Imai, Kenichiro, Nozaki, Tomoyoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465240/
https://www.ncbi.nlm.nih.gov/pubmed/32806678
http://dx.doi.org/10.3390/microorganisms8081229
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author Santos, Herbert J.
Chiba, Yoko
Makiuchi, Takashi
Arakawa, Saki
Murakami, Yoshitaka
Tomii, Kentaro
Imai, Kenichiro
Nozaki, Tomoyoshi
author_facet Santos, Herbert J.
Chiba, Yoko
Makiuchi, Takashi
Arakawa, Saki
Murakami, Yoshitaka
Tomii, Kentaro
Imai, Kenichiro
Nozaki, Tomoyoshi
author_sort Santos, Herbert J.
collection PubMed
description Mitochondrial matrix proteins synthesized in the cytosol often contain amino (N)-terminal targeting sequences (NTSs), or alternately internal targeting sequences (ITSs), which enable them to be properly translocated to the organelle. Such sequences are also required for proteins targeted to mitochondrion-related organelles (MROs) that are present in a few species of anaerobic eukaryotes. Similar to other MROs, the mitosomes of the human intestinal parasite Entamoeba histolytica are highly degenerate, because a majority of the components involved in various processes occurring in the canonical mitochondria are either missing or modified. As of yet, sulfate activation continues to be the only identified role of the relic mitochondria of Entamoeba. Mitosomes influence the parasitic nature of E. histolytica, as the downstream cytosolic products of sulfate activation have been reported to be essential in proliferation and encystation. Here, we investigated the position of the targeting sequence of one of the mitosomal matrix enzymes involved in the sulfate activation pathway, ATP sulfurylase (AS). We confirmed by immunofluorescence assay and subcellular fractionation that hemagluttinin (HA)-tagged EhAS was targeted to mitosomes. However, its ortholog in the δ-proteobacterium Desulfovibrio vulgaris, expressed as DvAS-HA in amoebic trophozoites, indicated cytosolic localization, suggesting a lack of recognizable mitosome targeting sequence in this protein. By expressing chimeric proteins containing swapped sequences between EhAS and DvAS in amoebic cells, we identified the ITSs responsible for mitosome targeting of EhAS. This observation is similar to other parasitic protozoans that harbor MROs, suggesting a convergent feature among various MROs in favoring ITS for the recognition and translocation of targeted proteins.
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spelling pubmed-74652402020-09-04 Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences Santos, Herbert J. Chiba, Yoko Makiuchi, Takashi Arakawa, Saki Murakami, Yoshitaka Tomii, Kentaro Imai, Kenichiro Nozaki, Tomoyoshi Microorganisms Article Mitochondrial matrix proteins synthesized in the cytosol often contain amino (N)-terminal targeting sequences (NTSs), or alternately internal targeting sequences (ITSs), which enable them to be properly translocated to the organelle. Such sequences are also required for proteins targeted to mitochondrion-related organelles (MROs) that are present in a few species of anaerobic eukaryotes. Similar to other MROs, the mitosomes of the human intestinal parasite Entamoeba histolytica are highly degenerate, because a majority of the components involved in various processes occurring in the canonical mitochondria are either missing or modified. As of yet, sulfate activation continues to be the only identified role of the relic mitochondria of Entamoeba. Mitosomes influence the parasitic nature of E. histolytica, as the downstream cytosolic products of sulfate activation have been reported to be essential in proliferation and encystation. Here, we investigated the position of the targeting sequence of one of the mitosomal matrix enzymes involved in the sulfate activation pathway, ATP sulfurylase (AS). We confirmed by immunofluorescence assay and subcellular fractionation that hemagluttinin (HA)-tagged EhAS was targeted to mitosomes. However, its ortholog in the δ-proteobacterium Desulfovibrio vulgaris, expressed as DvAS-HA in amoebic trophozoites, indicated cytosolic localization, suggesting a lack of recognizable mitosome targeting sequence in this protein. By expressing chimeric proteins containing swapped sequences between EhAS and DvAS in amoebic cells, we identified the ITSs responsible for mitosome targeting of EhAS. This observation is similar to other parasitic protozoans that harbor MROs, suggesting a convergent feature among various MROs in favoring ITS for the recognition and translocation of targeted proteins. MDPI 2020-08-12 /pmc/articles/PMC7465240/ /pubmed/32806678 http://dx.doi.org/10.3390/microorganisms8081229 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Santos, Herbert J.
Chiba, Yoko
Makiuchi, Takashi
Arakawa, Saki
Murakami, Yoshitaka
Tomii, Kentaro
Imai, Kenichiro
Nozaki, Tomoyoshi
Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title_full Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title_fullStr Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title_full_unstemmed Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title_short Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences
title_sort import of entamoeba histolytica mitosomal atp sulfurylase relies on internal targeting sequences
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465240/
https://www.ncbi.nlm.nih.gov/pubmed/32806678
http://dx.doi.org/10.3390/microorganisms8081229
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